The application of phosphate-solubilizing bacteria (PSB) has received little attention in aquaculture. In addition, the low efficiency of PSB as a biofertilizer in farm conditions is a major concern. Therefore, this study aims to isolate the PSB from sediment of earthen fishponds and evaluate with a more appropriate approach to ensure their effectiveness in increasing the bioavailability of phosphorus (P) in farm conditions. PSB was first isolated and selected using the National Botanical Research Institute's Phosphate (NBRIP) medium-containing tri-calcium phosphate (TCP) in solid and liquid media. Among 96 strains that were isolated, 11 strains identified by 16 s rRNA, belonging to the genera Pseudomonas and Acinetobacter, showed a higher ability to release P from TCP (48-170 mg L-1 ). Then, the efficiency of 11 strains was evaluated by combining different criteria. Among 11 selected strains, based on the ability to dissolve TCP and Ca-Phytate in culture medium, release P in sediment microcosm, and growth in a wide range of environmental conditions in fishponds, especially optimum growth at 4°C and pH above 8, Ps. deceptionensis strain Persian10 was selected as the most efficient strain for testing in aquarium conditions. In the last step, incubation of Persian10 in aquarium sediment (sterilized under gamma-ray) increased soluble P and had an impact on calcium phosphate, organic P and alkaline phosphatase activity; however, Persian10 had no impact on the concentration of iron phosphate and aluminium phosphate. Persian 10 strain can be considered a bio-fertilizer candidate in earthen fishponds as it expressed the ability to solubilize P in different conditions. It is very important to evaluate PSB in an appropriate process using a combination of different criteria to ensure optimal performance of PSB in farm conditions.

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